Gh secretagogues and uses thereof

ABSTRACT

The invention relates to use of a GH secretagogue (e.g. GRF or an analog thereof) for (1) altering a lipid parameter in a subject; (2) altering a body composition parameter in a subject, (3) treating a condition characterized by deficient or decreased bone formation in a subject (4) improving daytime vigilance and/or cognitive function in a subject, (5) improving a metabolic condition in a subject, (6) improving anabolism in a catabolic condition in a subject, and/or (7) improving and/or reconstituting immune function in a subject.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation in part of PCT International PatentApplication No. PCT/CA03/00827 filed May 29, 2003, which is incorporatedherein in its entirety.

FIELD OF THE INVENTION

The invention relates to growth hormone (GH) secretagogues, such asGH-releasing factor (GRF) and analogs thereof, and uses thereof.

BACKGROUND OF THE INVENTION Syndromes Associated with Fat Accumulation

Physiological systems include a number of lipid compounds/molecules suchas triglycerides and sterols (e.g. cholesterol), and associatedcarriers/complexes such as HDL, LDL, etc. Adverse levels or metabolismof such molecules is associated with related conditions such as fataccumulation, and associated disease.

Fat accumulation is observed in a range of conditions or syndromes suchas obesity, metabolic syndrome, and the recently described HIV-relatedlipodystrophy syndrome. All these conditions include features which areknown to increase the risk of diabetes and/or cardiovascular diseases.

The metabolic syndrome, also known as syndrome X, affect persons withfrank obesity as well as those with an increased amount of abdominalfat, and is characterized by insulin resistance, dyslipidemia(hypertriglyceridemia, low serum HDL cholesterol levels, and increasedLDL cholesterol levels) and hypertension.

HIV-infected patients treated with highly active antiretroviral therapy(HAART) commonly experience changes in fat distribution that includeincreased visceral and central fat accumulation (1), as well as loss ofextremity and subcutaneous fat (especially in the facial fat pads, limbsand buttocks) in association with insulin resistance and dyslipidemia(2, 3). Recent data suggest increased cardiovascular disease andmyocardial infarction rates in patients treated with prolongedantiretroviral therapy (ART) (4). In non HIV-infected patients (5) andamong HIV-infected patients with changes in fat distribution (6),increased waist to hip ratio (WHR) and central fat accumulation isrelated to increased metabolic risk indices.

Growth Hormone is known for its lipolytic properties, and its potentialrole in reversing several of the body fat and associated metabolicabnormalities has been actively studied. Beneficial effects have beenshown in GH-deficient individuals (Gotherstrom G. et al., J ClinEndocrinol Metab 2001 86(10):4657-4665) and non-HIV patients withabdominal obesity (Johannsson G, et al., J Clin Endocrinol Metab 1997,82(3):727-734). Recent studies also suggest that that GH levels arereduced in HIV-infected patients, and correlate inversely with excessvisceral fat accumulation (7, 8). Studies using higher dose,pharmacologic GH administration have resulted in reduced visceraladiposity in this population, but are associated with increased insulinresistance and side effects (9-12).

Cognitive Function

Cognitive abilities are impaired in a number of conditions includingadvancing age. Deleterious changes observed with aging affectparticularly fluid intelligence, or abilities involving conceptformation, rule discovery, planning behavior, and non-verbal reasoning.Conversely, crystallized intelligence, or abilities dependent uponaccumulated experience and education is relatively resistant toage-related decline. It has been suggested that the decline in GH andIGF-1 observed with aging contribute to the impaired cognitive function.

Evidence exists from both animal and human studies that administrationof GRF, GH or IGF-1 has significant effect on cognitive functions inconditions where these functions are impaired. For example, this hasbeen demonstrated with GH therapy in GH-deficient adults (Deijen J B, etal., Psychoneuroendocrinology 1998 23(1):45-55), and with administrationof IGF-1 or GRF in the healthy elderly (Aleman A et al., J ClinEndocrinol Metab 1999 84(2):471-475; Vitiello M. V., et al.,Gerontologist 2002 40(Special Issue 1):39).

Immune Function

Aging is accompanied by diminished circulating GH and IGF-1 levelsobserved in parallel with a declined function of the immune system,particularly affecting the T-cell mediated immunity. The age-relatedT-cell immune deficiency has been partly attributed to a progressiveatrophy of the thymus gland and is considered to be causally related tothe increased risk and severity of acquired infections observed in theelderly.

GH and IGF-1 are known to play an integrating role in the developmentand function of the immune system, as endocrine and/orautocrine/paracrine factors, and their administration has been shown toreverse age-related immune changes. Immune enhancing effects of thesefactors have been investigated in other immune deficiency states andencouraging results have been observed in HIV-positive patients(Napolitano L A, et al., AIDS 2002 16(8):1103-1111) and in animal modelsof radiotherapy preceding bone marrow transplantation (Sun R, et al.,BMT Meetings, February 22-26 Orlando, Fla., Abstract 27 2002:68-69).

Catabolism or Muscle Wasting

Muscle protein catabolism, or muscle wasting, accompanies many diseasesincluding all critical illness, regardless of the primary cause ofdisease. It is an important factor for the long-term prognosis and thelength of hospital stay and recovery, and may also be a limiting factorfor survival. Although many therapeutic tools have been investigatedincluding specific nutritional treatment, there is still a strong needfor more effective strategies to counteract protein catabolism.

Previous studies have reported that GH treatment increases muscle massin older patients. The anabolic effects or abilities of GH to reverse orattenuate muscle wasting have been investigated in several patientgroups. GH has been shown to improve nitrogen balance, an index of netwhole-body protein balance, after major gastro-intestinal surgery, burninjury, or major trauma. Anabolic effects have been translated intoclinical benefits in COPD patients (improvement of the maximalinspiratory pressure) (Papte G S, et al., Chest 1991 99(6):1495-1500)and elderly patient undergoing surgery following hip fracture(improvement of functional recovery defined as return to independence)(Van der Lely A J, et al., Eur J Endocrinol 2000 143(5):585-592).Finally, rGH has been recently approved for management of AIDS-wastingbased on results showing increased body weight, lean body mass andfunctional performance following 12 weeks of treatment (Schambelan M, etal., Ann Intern Med 1996 125(11):873-882).

However, the use of GH to treat conditions such as those noted above,has been associated with adverse side effects in some cases.

SUMMARY OF THE INVENTION

The invention relates to GH secretagogues (e.g. GRF and analogs thereof)and uses thereof.

Therefore, in a first aspect, the invention provides a method ofaltering a lipid parameter in a subject, said method comprisingadministering to said subject an agent selected from the groupconsisting of: (a) a growth hormone (GH) secretagogue; and (b) acomposition comprising a GH secretagogue and a pharmaceuticallyacceptable carrier.

In a further aspect, the invention provides a package comprising anagent selected from the group consisting of (a) a growth hormone (GH)secretagogue; and (b) a composition comprising a GH secretagogue and apharmaceutically acceptable carrier; together with instructions foraltering a lipid parameter in a subject.

In a further aspect, the invention provides a use of an agent selectedfrom the group consisting of (a) a growth hormone (GH) secretagogue; and(b) a composition comprising a GH secretagogue and a pharmaceuticallyacceptable carrier; for altering a lipid parameter in a subject.

In a further aspect, the invention provides a use of a growth hormone(GH) secretagogue for the preparation of a medicament for altering alipid parameter in a subject.

In a further aspect, the invention provides a method of altering a firstbody composition parameter of a subject, the method comprisingadministering to said subject an agent selected from the groupconsisting of (a) a growth hormone (GH) secretagogue; and (b) acomposition comprising a GH secretagogue and a pharmaceuticallyacceptable carrier.

In a further aspect, the invention provides a package comprising anagent selected from the group consisting of: (a) a growth hormone (GH)secretagogue; and (b) a composition comprising a GH secretagogue and apharmaceutically acceptable carrier; together with instructions foraltering a first body composition parameter of a subject.

In a further aspect, the invention provides a use of an agent selectedfrom the group consisting of: (a) a growth hormone (GH) secretagogue;and (b) a composition comprising a GH secretagogue and apharmaceutically acceptable carrier; for altering a first bodycomposition-parameter of a subject.

In a further aspect, the invention provides a use of a growth hormone(GH) secretagogue for the preparation of a medicament for altering afirst body composition parameter of a subject.

In a further aspect, the invention provides a method of treating acondition characterized by deficient or decreased bone formation in asubject, said method comprising administering to said subject an agentselected from the group consisting of: (a) a growth hormone (GH)secretagogue; and (b) a composition comprising a GH secretagogue and apharmaceutically acceptable carrier.

In a further aspect, the invention provides a package comprising anagent selected from the group consisting of: (a) a growth hormone (GH)secretagogue; and (b) a composition comprising a GH secretagogue and apharmaceutically acceptable carrier; together with instructions fortreating a condition characterized by deficient or decreased boneformation in a subject.

In a further aspect, the invention provides a use of an agent selectedfrom the group consisting of: (a) a growth hormone (GH) secretagogue;and (b) a composition comprising a GH secretagogue and apharmaceutically acceptable carrier; for treating a conditioncharacterized by deficient or decreased bone formation in a subject.

In a further aspect, the invention provides a use of a growth hormone(GH) secretagogue for the preparation of a medicament for treating acondition characterized by deficient or decreased bone formation in asubject.

In further aspects, the invention relates to a method for (1)stimulating day-time vigilance and/or cognitive functions e.g. inconditions related to aging or mild cognitive impairment, (2) improvingmetabolic conditions associated with fat accumulation and/orhypercholesterolemia, (e.g. metabolic conditions including obesity,HIV-related lipodystrophy, metabolic syndrome or syndrome X), (3)improving anabolism in catabolic/wasting conditions (such as thoseobserved in Chronic Renal Failure, congestive heart failure AIDS,following hip fracture, trauma, or major surgery, particularly inelderly subjects), and/or (4) improving immune function orreconstitution of immunodeficient states such as that associated aging,HIV or following high-dose chemotherapy and/or radiotherapy; the methodcomprising administering a GH secretagogue (e.g. GRF and analogsthereof) or a composition comprising a GH secretagogue and apharmaceutically acceptable carrier; to a subject.

In further aspects, the invention relates to uses of a GH secretagogue(e.g. GRF and analogs thereof) or a composition comprising a GHsecretagogue and a pharmaceutically acceptable carrier, for (1)stimulating day-time vigilance and/or cognitive functions e.g. inconditions related to aging or mild cognitive impairment, (2) improvingmetabolic conditions associated with fat accumulation and/orhypercholesterolemia, (e.g. metabolic conditions including obesity,HIV-related lipodystrophy, metabolic syndrome or syndrome X), (3)improving anabolism in catabolic/wasting conditions (such as thoseobserved in Chronic Renal Failure, congestive heart failure AIDS,following hip fracture, trauma, or major surgery, particularly inelderly subjects), and/or (4) improving immune function orreconstitution of immunodeficient states such as that associated aging,HIV or following high-dose chemotherapy and/or radiotherapy.

In further aspects, the invention similarly relates to a packagecomprising an agent selected from the group consisting of: (a) a growthhormone (GH) secretagogue; and (b) a composition comprising a GHsecretagogue and a pharmaceutically acceptable carrier; together withinstructions for (1) stimulating day-time vigilance and/or cognitivefunctions (2) improving metabolic conditions associated with fataccumulation and/or hypercholesterolemia, (3) improving anabolism incatabolic/wasting conditions, and/or (4) improving immune function orreconstitution of immunodeficient states.

In further aspects, the invention similarly relates to a use of a GHsecretagogue for the preparation of a medicament for (1) stimulatingday-time vigilance and/or cognitive functions (2) improving metabolicconditions associated with fat accumulation and/or hypercholesterolemia,(3) improving anabolism in catabolic/wasting conditions, and/or (4)improving immune function or reconstitution of immunodeficient states.

In accordance with the present invention, there is provided acomposition for (1) altering a lipid parameter in a subject; (2)altering a body composition parameter in a subject, (3) treating acondition characterized by deficient or decreased bone formation in asubject (4) improving daytime vigilance and/or cognitive function in asubject, (5) improving a metabolic condition in a subject, (6) improvinganabolism in a catabolic condition in a subject, and/or (7) improvingand/or reconstituting immune function in a subject, the compositioncomprising an effective amount of a GH secretagogue (e.g. GRF compoundor analog thereof) in association with a pharmaceutically acceptablecarrier, excipient or diluent.

In embodiments, the metabolic condition is associated with fataccumulation and/or hypercholesterolemia, e.g. obesity, HIV-relatedlipodystrophy, metabolic syndrome and syndrome X.

In embodiments the catabolic condition is related to one selected fromthe group consisting of chronic renal failure, AIDS, hip fracture,trauma or major surgery in a subject.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates the differences between treatment groups in changesfrom baseline to week 2 in the mean reaction time of the ContinuousPerformance Test (CPT);

FIG. 2 illustrates changes from baseline to day 9 in Pz amplitude ofevoked related potential (P300) during wakefulness;

FIG. 3 illustrates mean AUC of antigen-specific proliferative T cellresponse;

FIG. 4 illustrates the percentage of subjects with protective antibodytiters (> 1/40) for B/Victoria; and

FIG. 5 illustrates the variation of mean IGF-1 levels during time withplacebo, 2 mg/day TH9507, 0.5 mg/day TH9507 and 1 mg/day TH9507.

FIG. 6 illustrates a flow diagram of patient disposition in respect ofthe studies described in Example 6.

FIG. 7 illustrates results of dose response of TH9507 on A) IGF-I and B)truncal fat, as measured by DEXA, as described in Example 6. 7A: “ng/ml”represents ng/ml of TGF-1 as measured. 7B: “kg” represents change intruncal fat, with the negative values indicating a decrease in truncalfat. 7A and 7B: “placebo”, “1 mg” and “2 mg” correspond toadministration of a placebo, 1 mg of Th9507 and 2 mg of Th9507,respectively. Results are mean (SD). *=P<0.01 vs. placebo by ANOVA.

DETAILED DESCRIPTION OF THE INVENTION

In the studies described herein, the effect of (hexenoyltrans-3)hGRF(1-44)NH₂ (also referred to as TH9507 herein), a growthhormone releasing factor (GRF) analog, was assessed over 12 weeks inHIV-infected men and women with evidence of fat redistribution andincreased truncal adiposity. The data herein demonstrate significanteffects of TH9507 to increase lean body mass and reduce truncal fat andvisceral fat and improve lipid parameters, sparing extremity andsubcutaneous fat (Example 6). These effects were achieved withphysiological increases in GH secretion and without adverse effects onblood glucose levels.

The studies described herein further demonstrate beneficial effects onbone markers.

The studies described herein further demonstrate a beneficial effect ofTh9507 on daytime vigilance and cognitive function (Example 2), immuneresponse (Example 3), wasting/catabolic conditions (Example 4) andnon-HDL cholesterol (Example 5).

Accordingly, in a first aspect, the invention provides a method ofaltering a lipid parameter in a subject, said method comprisingadministering to said subject an agent selected from the groupconsisting of: (a) a growth hormone (GH) secretagogue; and (b) acomposition comprising a GH secretagogue and a pharmaceuticallyacceptable carrier. In an embodiment, the method results in no orsubstantially no increase in blood glucose levels in said subject.

In a further aspect, the invention provides a package comprising anagent selected from the group consisting of (a) a growth hormone (GH)secretagogue; and (b) a composition comprising a GH secretagogue and apharmaceutically acceptable carrier; together with instructions foraltering a lipid parameter in a subject. In an embodiment, theabove-mentioned alteration of a lipid parameter results in no orsubstantially no increase in blood glucose levels in said subject.

In a further aspect, the invention provides a use of an agent selectedfrom the group consisting of (a) a growth hormone (GH) secretagogue; and(b) a composition comprising a GH secretagogue and a pharmaceuticallyacceptable carrier; for altering a lipid parameter in a subject. In anembodiment, the above-mentioned alteration of a lipid parameter resultsin no or substantially no increase in blood glucose levels in saidsubject.

In a further aspect, the invention provides a use of a growth hormone(GH) secretagogue for the preparation of a medicament for altering alipid parameter in a subject. In an embodiment, the above-mentionedalteration of a lipid parameter results in no or substantially noincrease in blood glucose levels in said subject.

In an embodiment, the above-noted the alteration of a lipid parameter isselected from the group consisting of: (a) a decrease in cholesterol;(b) a decrease in non-HDL cholesterol; (c) a decrease in triglyceride;(d) a decrease in the ratio of total cholesterol:HDL cholesterol; and(e) any combination of (a) to (d).

In an embodiment, the above-noted lipid parameter is associated with acondition selected from the group consisting of lipodystrophy,lipohypertrophy, obesity, dyslipidemia, hypertriglyceridemia andsyndrome X.

In a further aspect, the invention provides a method of altering a firstbody composition parameter of a subject, the method comprisingadministering to said subject an agent selected from the groupconsisting of (a) a growth hormone (GH) secretagogue; and (b) acomposition comprising a GH secretagogue and a pharmaceuticallyacceptable carrier. In an embodiment, the method results in no orsubstantially no decrease in a second body composition parameter of thesubject, wherein the second body composition parameter is selected fromthe group consisting of: (i) limb fat; (ii) subcutaneous fat; (iii)subcutaneous abdominal tissue (SAT); and (iv) any combination of (i) to(iii).

In a further aspect, the invention provides a package comprising anagent selected from the group consisting of: (a) a growth hormone (GH)secretagogue; and (b) a composition comprising a GH secretagogue and apharmaceutically acceptable carrier; together with instructions foraltering a first body composition parameter of a subject. In anembodiment, the above-mentioned alteration of a first body compositionparameter results in no or substantially no decrease in a second bodycomposition parameter of the subject, wherein the second bodycomposition parameter is selected from the group consisting of (i) limbfat; (ii) subcutaneous fat; (iii) subcutaneous abdominal tissue (SAT);and (iv) any combination of (i) to (iii).

In a further aspect, the invention provides a use of an agent selectedfrom the group consisting of: (a) a growth hormone (GH) secretagogue;and (b) a composition comprising a GH secretagogue and apharmaceutically acceptable carrier; for altering a first bodycomposition parameter of a subject. In an embodiment, theabove-mentioned alteration of a first body composition parameter resultsin no or substantially no decrease in a second body compositionparameter of the subject, wherein the second body composition parameteris selected from the group consisting of (i) limb fat; (ii) subcutaneousfat; (iii) subcutaneous abdominal tissue (SAT); and (iv) any combinationof (i) to (iii).

In a further aspect, the invention provides a use of a growth hormone(GH) secretagogue for the preparation of a medicament for altering afirst body composition parameter of a subject. In an embodiment, theabove-mentioned alteration of a first body composition parameter resultsin no or substantially no decrease in a second body compositionparameter of the subject, wherein the second body composition parameteris selected from the group consisting of (i) limb fat; (ii) subcutaneousfat; (iii) subcutaneous abdominal tissue (SAT); and (iv) any combinationof (i) to (iii).

In an embodiment, the above-mentioned alteration of a first bodycomposition parameter is selected from the group consisting of: (a) anincrease in lean body mass; (b) a decrease in trunk fat; (c) a decreasein visceral fat; (d) a decrease in abdominal girth; (e) a decrease invisceral abdominal tissue (VAT); (f) a decrease in VAT:SAT ratio; and(g) any combination of (a) to (f).

In an embodiment, the first body composition parameter is associatedwith a condition selected from the group consisting of lipodystrophy,lipohypertrophy, obesity, dyslipidemia, hypertriglyceridemia andsyndrome X.

In an embodiment, the above-mentioned alteration of a first bodycomposition parameter results in an improvement of quality of life ofthe subject.

In a further aspect, the invention provides a method of treating acondition characterized by deficient or decreased bone formation or fortreating bone dysfunction or defect, in a subject, said methodcomprising administering to said subject an agent selected from thegroup consisting of: (a) a growth hormone (GH) secretagogue; and (b) acomposition comprising a GH secretagogue and a pharmaceuticallyacceptable carrier.

In a further aspect, the invention provides a package comprising anagent selected from the group consisting of: (a) a growth hormone (GH)secretagogue; and (b) a composition comprising a GH secretagogue and apharmaceutically acceptable carrier; together with instructions fortreating a condition characterized by deficient or decreased boneformation in a subject.

In a further aspect, the invention provides a use of an agent selectedfrom the group consisting of: (a) a growth hormone (GH) secretagogue;and (b) a composition comprising a GH secretagogue and apharmaceutically acceptable carrier; for treating a conditioncharacterized by deficient or decreased bone formation in a subject.

In a further aspect, the invention provides a use of a growth hormone(GH) secretagogue for the preparation of a medicament for treating acondition characterized by deficient or decreased bone formation in asubject.

In an embodiment, the above-noted condition characterized by deficientor decreased bone formation is selected from the group consisting ofosteopenia and osteoporosis.

In further aspects, the invention relates to uses of a GH secretagogue(e.g. GRF and analogs thereof) or a composition comprising a GHsecretagogue and a pharmaceutically acceptable carrier, for (1)stimulating day-time vigilance and/or cognitive functions e.g. inconditions related to aging or mild cognitive impairment, (2) improvingmetabolic conditions associated with fat accumulation and/orhypercholesterolemia, (e.g. metabolic conditions including obesity,HIV-related lipodystrophy, metabolic syndrome or syndrome X), (3)improving anabolism in catabolic/wasting conditions (such as thoseobserved in Chronic Renal Failure, congestive heart failure AIDS,following hip fracture, trauma, or major surgery, particularly inelderly subjects), and/or (4) improving immune function orreconstitution of immunodeficient states such as that associated aging,HIV or following high-dose chemotherapy and/or radiotherapy.

In further aspects, the invention similarly relates to a packagecomprising an agent selected from the group consisting of: (a) a growthhormone (GH) secretagogue; and (b) a composition comprising a GHsecretagogue and a pharmaceutically acceptable carrier; together withinstructions for (1) stimulating day-time vigilance and/or cognitivefunctions (2) improving metabolic conditions associated with fataccumulation and/or hypercholesterolemia, (3) improving anabolism incatabolic/wasting conditions, and/or (4) improving immune function orreconstitution of immunodeficient states.

In further aspects, the invention similarly relates to a use of a GHsecretagogue for the preparation of a medicament for (1) stimulatingday-time vigilance and/or cognitive functions (2) improving metabolicconditions associated with fat accumulation and/or hypercholesterolemia,(3) improving anabolism in catabolic/wasting conditions, and/or (4)improving immune function or reconstitution of immunodeficient states.

In an embodiment, the above-mentioned lipodystrophy is HIV-relatedlipodystrophy.

In an embodiment, the above-mentioned subject is HIV positive.

In an embodiment, the above-mentioned subject is receiving antiviraltherapy.

In an embodiment, the above-mentioned subject suffers from a conditionselected from the group consisting of diabetes, glucose intolerance andinsulin resistance.

In an embodiment the above-mentioned GH secretagogue is administered ata dose of about 0.0001 to about 4 mg, in a further embodiment, about0.0001 to about 2 mg, in a further embodiment, about 1 mg to about 2 mg,in a further embodiment, about 1 mg and in a further embodiment, about 2mg.

In an embodiment, the agent is administered by a route selected from thegroup consisting of intravenous, oral, transdermal, subcutaneous,mucosal, intramuscular, intranasal, intrapulmonary, parenteral,intrarectal and topical. In an embodiment, the agent is administered bya subcutaneous route.

In an embodiment, the subject is a mammal, in a further embodiment, ahuman.

For the purpose of the present invention the following terms are definedbellow.

The term “analog” is intended to mean a molecule of different structurebut having a biological function similar to the structures of the GRF orto a biologically functional fragment thereof which may includepeptidomimetics. Peptidomimetics may be conveniently prepared by directchemical synthesis using methods well known in the art.

The term “subject” is intended to mean any mammal including, but notlimited to, human, canine, feline, equine, caprine, bovine, porcine andovine.

The term “cognitive function” is intended to mean functions including,but not limited to thinking, reasoning, memory and problem solving.

The term “catabolic/wasting conditions” is intended to mean conditionincluding, but not limited to, frail bones, low muscular mass and musclewasting.

In the present application, the compound identified as TH9507 is the[hexenoyl-trans-3-Tyr1]hGRF(1-44)NH₂.

In various aspects, the methods and corresponding uses and packagesdescribed herein comprise administering to a subject a growth hormone(GH) secretagogue. “GH secretagogue” as used herein refers to anycompound or molecule, natural or synthetic, which may result in, eitherdirectly or indirectly, GH secretion and/or an increase in GH secretion.

In embodiments, the GH secretagogue is a growth hormone-releasing factor(GRF; also referred to as growth hormone releasing hormone [GHRH]) orGRF analog.

In an embodiment, the GRF is human GRF (hGRF).

Human growth hormone-releasing factor (hGRF) is a peptide of 44 aminoacids with a C-terminal NH₂ modification, referred to herein ashGRF(1-44)NH₂, and has the following structure:

(SEQ ID NO: 2) Tyr-Ala-Asp-Ala-Ile-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-Gln-Leu-Ser-Ala-Arg-Lys-Leu-Leu-Gln-Asp-Ile-Met-Ser-Arg-Gln-Gln-Gly-Glu-Ser-Asn-Gln-Glu-Arg-Gly-Ala-Arg-Ala-Arg-Leu-NH₂

Therefore, the amino acid sequence of the just-noted 44 amino acid formis as follows:

(SEQ ID NO: 3) Tyr-Ala-Asp-Ala-Ile-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-Gln-Leu-Ser-Ala-Arg-Lys-Leu-Leu-Gln-Asp-Ile-Met-Ser-Arg-Gln-Gln-Gly-Glu-Ser-Asn-Gln-Glu-Arg-Gly-Ala-Arg-Ala-Arg-Leu

The minimum active core comprises the first 29 amino acids of the abovesequence, which is referred to herein as hGRF(1-29)NH₂, and has thefollowing structure:

(SEQ ID NO: 4) Tyr-Ala-Asp-Ala-Ile-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-Gln-Leu-Ser-Ala-Arg-Lys-Leu-Leu-Gln- Asp-Ile-Met-Ser-Arg-NH₂

Therefore, the amino acid sequence of the just-noted 29 amino acid formis as follows:

(SEQ ID NO: 5) Tyr-Ala-Asp-Ala-Ile-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-Gln-Leu-Ser-Ala-Arg-Lys-Leu-Leu-Gln- Asp-Ile-Met-Ser-Arg

The 1-44 and 1-29 forms differ in that the 1-44 form contains thefollowing additional amino acids, which correspond to positions 30-44 ofthe 1-44 form:

(SEQ ID NO: 6) Gln-Gln-Gly-Glu-Ser-Asn-Gln-Glu-Arg-Gly-Ala-Arg-Ala-Arg-Leu

In an embodiment, the above-mentioned GRF analog is a GRF analog offormula A:

X-GRF Peptide (A)

-   -   wherein;    -   the GRF peptide is a peptide of formula B;

(SEQ ID NO: 1) A1-A2-Asp-Ala-Ile-Phe-Thr-A8-Ser-Tyr-Arg-Lys-A13-Leu-A15-Gln-Leu-A18-Ala-Arg-Lys-Leu-Leu-A24-A25- Ile-A27-A28-Arg-A30-R0(B)

-   -   wherein,        -   A1 is Tyr or His;        -   A2 is Val or Ala;        -   A8 is Asn or Ser;        -   A13 is Val or Ile;        -   A15 is Ala or Gly;        -   A18 is Ser or Tyr;        -   A24 is Gln or His;        -   A25 is Asp or Glu;        -   A27 is Met, Ile or Nle;        -   A28 is Ser or Asn;        -   A30 is a bond or amino acid sequence of 1 up to 15 residues;            and        -   R0 is NH₂ or NH—(CH₂)n-CONH₂, with n=1 to 12; and    -   X is a hydrophobic tail anchored via an amide bond to the        N-terminus of the peptide and the hydrophobic tail defining a        backbone of 5 to 7 atoms;    -   wherein the backbone can be substituted by C1-6 alkyl, C3-6        cycloalkyl, or C6-12 aryl and the backbone comprises at least        one rigidifying moiety connected to at least two atoms of the        backbone;    -   said moiety selected from the group consisting of double bond,        triple bond, saturated or unsaturated C3-9 cycloalkyl, and C6-12        aryl.

In embodiments, X noted above is selected from the group consisting of:

In embodiments, A30 noted above is selected from the group consistingof:

-   -   (a) a bond;    -   (b) an amino acid sequence corresponding to positions 30-44 of a        natural GRF peptide, and    -   (c) said amino acid sequence of (b) having a 1-14 amino acid        deletion from its C-terminal.

In embodiments, the above-noted GRF peptide is selected from the groupconsisting of:

-   -   (a) a polypeptide comprising the amino acid sequence of SEQ ID        NO: 3;    -   (b) a polypeptide comprising the amino acid sequence of SEQ ID        NO: 5; and    -   (c) the polypeptide of (a) having a 1 to 14 amino acid deletion        from its C-terminus.

In an embodiment, the above-noted GRF analog is (hexenoyltrans-3)hGRF(1-44)NH₂.

Methods of preparing the above-described GRF analogs are described inU.S. Pat. No. 5,861,379 (Ibea et al., Jan. 19, 1999); U.S. Pat. No.6,020,311 (Brazeau et al., Feb. 1, 2000), U.S. Pat. No. 6,458,764(Gravel et al., Oct. 1, 2002) and published US application No.2004/0171534 A1 (Gravel et al., published Sep. 2, 2004).

As noted above, in various embodiments, the above-mentioned GHsecretagogue may be used therapeutically in formulations or medicamentsto effect the above-noted alterations and to prevent or treat theabove-noted conditions. The invention provides corresponding methods ofmedical treatment, in which a therapeutic dose of a GH secretagogue isadministered in a pharmacologically acceptable formulation, e.g. to apatient or subject in need thereof. Accordingly, the invention alsoprovides therapeutic compositions comprising a GH secretagogue and apharmacologically acceptable excipient or carrier. In one embodiment,such compositions include GH secretagogue in a therapeutically orprophylactically effective amount sufficient to effect the above-notedalterations and to treat the above-noted conditions. The therapeuticcomposition may be soluble in an aqueous solution at a physiologicallyacceptable pH.

A “therapeutically effective amount” refers to an amount effective, atdosages and for periods of time necessary, to achieve the desiredtherapeutic result, such as to effect the above-noted alterations and toreduce the progression of the above-noted conditions. A therapeuticallyeffective amount of a GH secretagogue may vary according to factors suchas the disease state, age, sex, and weight of the individual, and theability of the compound to elicit a desired response in the individual.Dosage regimens may be adjusted to provide the optimum therapeuticresponse. A therapeutically effective amount is also one in which anytoxic or detrimental effects of the compound are outweighed by thetherapeutically beneficial effects. A “prophylactically effectiveamount” refers to an amount effective, at dosages and for periods oftime necessary, to achieve the desired prophylactic result, such aspreventing or inhibiting the rate of onset or progression of theabove-noted conditions. A prophylactically effective amount can bedetermined as described above for the therapeutically effective amount.For any particular subject, specific dosage regimens may be adjustedover time according to the individual need and the professionaljudgement of the person administering or supervising the administrationof the compositions.

As used herein “pharmaceutically acceptable carrier” or “excipient”includes any and all solvents, dispersion media, coatings, antibacterialand antifungal agents, isotonic and absorption delaying agents, and thelike that are physiologically compatible. In one embodiment, the carrieris suitable for parenteral administration. Alternatively, the carriercan be suitable for intravenous, intraperitoneal, intramuscular,subcutaneous, sublingual or oral administration. Pharmaceuticallyacceptable carriers include sterile aqueous solutions or dispersions andsterile powders for the extemporaneous preparation of sterile injectablesolutions or dispersion. The use of such media and agents forpharmaceutically active substances is well known in the art. Exceptinsofar as any conventional media or agent is incompatible with theactive compound, use thereof in the pharmaceutical compositions of theinvention is contemplated. Supplementary active compounds can also beincorporated into the compositions.

Therapeutic compositions typically must be sterile and stable under theconditions of manufacture and storage. The composition can be formulatedas a solution, microemulsion, liposome, or other ordered structuresuitable to high drug concentration. The carrier can be a solvent ordispersion medium containing, for example, water, ethanol, polyol (forexample, glycerol, propylene glycol, and liquid polyethylene glycol, andthe like), and suitable mixtures thereof. The proper fluidity can bemaintained, for example, by the use of a coating such as lecithin, bythe maintenance of the required particle size in the case of dispersionand by the use of surfactants. In many cases, it will be preferable toinclude isotonic agents, for example, sugars, polyalcohols such asmannitol, sorbitol, or sodium chloride in the composition. Prolongedabsorption of the injectable compositions can be brought about byincluding in the composition an agent which delays absorption, forexample, monostearate salts and gelatin. Moreover, a GH secretagogue canbe administered in a time release formulation, for example in acomposition which includes a slow release polymer. The active compoundscan be prepared with carriers that will protect the compound againstrapid release, such as a controlled release formulation, includingimplants and microencapsulated delivery systems. Biodegradable,biocompatible polymers can be used, such as ethylene vinyl acetate,polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylacticacid and polylactic, polyglycolic copolymers (PLG). Many methods for thepreparation of such formulations are patented or generally known tothose skilled in the art.

Sterile injectable solutions can be prepared by incorporating the activecompound (e.g. a GH secretagogue ) in the required amount in anappropriate solvent with one or a combination of ingredients enumeratedabove, as required, followed by filtered sterilization. Generally,dispersions are prepared by incorporating the active compound into asterile vehicle which contains a basic dispersion medium and therequired other ingredients from those enumerated above. In the case ofsterile powders for the preparation of sterile injectable solutions, thepreferred methods of preparation are vacuum drying and freeze-dryingwhich yields a powder of the active ingredient plus any additionaldesired ingredient from a previously sterile-filtered solution thereof.In accordance with an alternative aspect of the invention, a GHsecretagogue may be formulated with one or more additional compoundsthat enhance its solubility.

In accordance with another aspect of the invention, therapeuticcompositions of the present invention, comprising a GH secretagogue, maybe provided in containers, kits or packages (e.g. commercial packages)which further comprise instructions for its use to effect theabove-noted alterations and to prevent or treat the above-notedconditions.

Accordingly, the invention further provides a package comprising a GHsecretagogue or the above-mentioned composition together withinstructions to effect the above-noted alterations and to prevent ortreat the above-noted conditions.

The invention further provides a use of a GH secretagogue to effect theabove-noted alterations and to prevent or treat the above-notedconditions. The invention further provides a use of a GH secretagoguefor the preparation of a medicament to effect the above-notedalterations and to prevent or treat the above-noted conditions.

Although various embodiments of the invention are disclosed herein, manyadaptations and modifications may be made within the scope of theinvention in accordance with the common general knowledge of thoseskilled in this art. Such modifications include the substitution ofknown equivalents for any aspect of the invention in order to achievethe same result in substantially the same way. Numeric ranges areinclusive of the numbers defining the range. In the claims, the word“comprising” is used as an open-ended term, substantially equivalent tothe phrase “including, but not limited to”. The following examples areillustrative of various aspects of the invention, and do not limit thebroad aspects of the invention as disclosed herein.

Throughout this application, various references are referred to describemore fully the state of the art to which this invention pertains. Thedisclosures of these references are hereby incorporated by referenceinto the present disclosure.

EXAMPLES Example 1 Study Drug

The compound used in the studies below is (hexenoyltrans-3)hGRF(1-44)NH₂ (also referred to as TH9507 herein), which is asynthetic human growth hormone releasing factor analog that comprisesthe 44-amino acid sequence of human growth hormone releasing factor(hGRF) on which a hexenoyl moiety, a C6 side chain has been anchored onTyr 1 at the n-terminal. (hexenoyl trans-3)hGRF(1-44)NH₂ or Th9507 hasthe following structure:

(SEQ ID NO: 7) (trans)CH₃—CH₂—CH═CH—CH₂—CO-Tyr-Ala-Asp-Ala-Ile-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-Gln-Leu-Ser-Ala-Arg-Lys-Leu-Leu-Gln-Asp-Ile-Met-Ser-Arg-Gln-Gln-Gly-Glu-Ser-Asn-Gln-Glu-Arg-Gly-Ala-Arg- Ala-Arg-Leu-NH₂(hexenoyl trans-3)hGRF(1-44)NH₂ was synthesized via the methods setforth in U.S. Pat. No. 5,861,379 (Ibea et al.; Jan. 19, 1999).

The in vitro half-life of Th9507 is 3-8 hours compared to 0.56 hours forhGRF. Daily 1 and 2 mg doses have been shown to increase IGF-I to thephysiological range seen in younger adults (14). The safety profile ofTH9507 is generally good. Th9507 has been studied in patients with TypeII DM, and was not shown to aggravate overall glycemic control whenadministered at a daily dose up to 2 mg (15).

Example 2 Administration of TH9507 for Improving Daytime Vigilance inSubjects with Sleep Maintenance Insomnia

The present example shows the effect of a 14 day-administration of 2doses of TH9507 (0.1 mg and 1 mg) on vigilance parameters in subjects of35 to 50 years of age exhibiting sleep maintenance insomnia.

Material and Methods

The study involved 82 patients exhibiting sleep maintenance insomnia (20females, 62 males; mean age 43.2±5.4 years). Patients were selectedbased on the Pittsburgh Sleep Quality Index (Score ≧5), the WaltersCriteria for Sleep Maintenance Insomnia, and the Beck Questionnaire(Score ≦17). The primary exclusion criteria were other primary sleepdisorders and the use of any products affecting sleep or vigilance inthe 30 days prior to entering the study.

The study was a randomized, double-blind, placebo-controlled, parallelgroup and multicenter evaluation of two doses of TH9507 (0.1 mg and 1mg) administered daily by subcutaneous injection at bedtime for 14consecutive days. To evaluate vigilance and the performance in themorning, patients underwent a battery of cognitive tests including theContinuous Performance Test (CPT) at baseline and at the end oftreatment period.

The CPT has been described in the literature as a measure of consistencyin responding and ability to sustain attention over time (Aleman A, etal., J Clin Endocrinol Metab 1999 84(2):471-475). This test requiredsubjects to press space bar each time the letter “A” was followed by“X”. Omission and commission errors, and Mean Reaction Time of correctresponses were analyzed.

Results

Demographic characteristics by treatment group are displayed in thefollowing table:

TABLE 1 Demographic (screening) data by treatment groups Placebo N = 290.1 mg N = 26 1 mg N = 27 P value Age (Mean ± SD) 44.0 ± 5.8  43.2 ±5.8  42.3 ± 4.5  0.53 (range) (35-56) (34-60) (34-50) (F-test) GenderMales 21 19 22 0.68 Females  8  7  5 chi-square test) Weight (kg) 0.94(Mean ± SD) 78.0 ± 14.3 78.2 ± 12.4 79.3 ± 15.3 (F-test)

As illustrated in FIG. 1, the Mean Reaction Time of the CPT wassignificantly and markedly decreased in the 0.1 mg group when comparedto placebo. The decrease from baseline to day 9 was 45.85 ms (P=0.023when compared to placebo as analyzed by an ANOVA model). No significanteffect was observed in the 1 mg group. Circulating IGF-1 and IGFBP-3levels were significantly increased at week 2 in the 1 mg group whencompared to placebo (P<0.0001, ANOVA on changes from baseline). Asexpected the 0.1 mg did not affect these parameters (P=0.07 and P=0.99)for IGF-1 and IGF-BP3 respectively, ANOVA model on changes frombaseline).

Additional data on effects of TH9507 on vigilance obtained in a previousstudy are presented in FIG. 2. In this study, TH9507 was administereddaily for 7 days in a cross-over design. This study involved 12 healthysubjects aged between 50 to 65 years, exhibiting age-related sleepimpairment (Pittsburgh Sleep Quality Index Score from 3 to 7). At theend of the treatment period, daytime vigilance was significantlyenhanced when compared to placebo in subjects receiving 1 mg of TH9507,as assessed by P300, an event-related potential test. Changes fromBaseline to day 9 in the Pz amplitude of the Evoked Related Potential(P300) observed in the placebo and 1 mg group were as follows: Placebo,−15%; 1 mg, +55%, P=0.0114, as analyzed by an ANOVA model.

In both studies, the safety profile of TH9507 was comparable to that ofthe placebo, except for a higher incidence of reactions at the site ofinjection observed at 1 mg in the insomnia study.

In summary, these results provide evidence that TH9507 improves daytimevigilance in sleep maintenance insomnia in subjects and would favor adirect mechanism of action of TH9507, not mediated by IGF-1. Data aresupported by those obtained by Vitiello et al (Vitiello M. V., et al.,Gerontologist 2002 40 (Special Issue 1):39-N/A. using hGRF in cognitivetests involving psychomotor and perceptual processing speed (Deijin J Bet al, Psychoneuroendocrinology 1998 23(1):45-55) and may supportfurther clinical investigations in subjects with impaired cognitivefunctions.

Example 3 Effects of TH9507 on the Immune Response to InfluenzaVaccination in Elderly Subjects

The present example describes immune findings following an influenzavaccination challenge in elderly subjects.

Material and Methods

Eighty seven (87) subjects aged 75 years in average were included in adouble-bind, randomized, placebo-controlled study. TH9507 or a placebowas administered at a daily dose of 1 or 2 mg by subcutaneous injectionfor 8 weeks. Follow-up assessments were conducted for 12 weeks after theend of the treatment period. At week 4, in the middle of the treatmentperiod, subjects received the commercial Canadian influenza vaccine(Vaccine Fluviral® S/F, Shire, Montreal, Canada) containing 15 μg eachof A/New Caledonia/20/99, A/Panama/2007/99, B/Victoria/504/2000antigens.

Influenza-specific proliferative T cell response and antibody titerswere evaluated for each of the 3 strains contained in the vaccine. Theproliferative T cell response was assessed by a mitogen assay usingtritiated thymidine (³H) incorporation and results were log-base 10transformed prior to analysis. The antibody titers were determined bystandard hemaglutinaion inhibition assay and results were log-base 2transformed prior to analysis.

Eighty one (81) subjects completed the study as per protocol. Subjectdemographics are shown in the following table:

TABLE 2 Subject demographics 1 mg 2 mg Placebo TH9507 TH9507 All P-valueAge 75.9 ± 6.5 74.9 ± 6.1 73.2 ± 4.4 74.6 ± 5.8 0.21 (years) Total N 2929 29 187  0.96 Female 13 13 14 40 Male 16 16 15 47 BMI 27.4 ± 5.8 26.9± 4.2 29.2 ± 6.0 27.8 ± 5.4 0.26 (kg/m²) Data for age and BMI arepresented as mean ± SD. Baseline comparability among treatment groupswas tested by ANOVA (age, BMI) or Pearson's chi-square test (gender).

Results

As shown in FIG. 3 the mean AUC calculated for the whole study periodincluding both the treatment and follow-up period (week 0 to week 20)was statistically higher in the 2 mg group when compared with placebo(Panama, P=0.03; New Caledonia, P=0.001; Victoria, P=0.02, (Pairwisecomparisons for difference among treatment groups and ANCOVA analysisfor overall treatment significance).

As illustrated in FIG. 4, administration of TH9507 increased theproportion of patients achieving a protective antibody level for theVictoria antigen when compared to placebo. This observation reachedstatistical significance at the 2 mg dose and was noted during both thetreatment and the follow-up periods (week 6: P=0.02; Week 8: P=0.01;week 12: P=0.02; week 16; P=0.004, week 20: P=0.01, pairwise comparisonsfor difference among treatment groups and Pearson chi-square test foroverall treatment difference) indicating a sustained effect for up to 16weeks after cessation of treatment. No statistical difference in thepercentage of subjects was observed in the Panama and New Caledoniastrains.

A dose-related increase in the mean IGF-1 values was observed during thewhole treatment period in both Th9507 groups when compared to baseline.Values returned to baseline following cessation of treatment.

No major difference in the incidence of adverse events was observedbetween treatment groups except for a dose-related trend in theincidence of reactions at the site of injection.

In summary, the findings observed in this study strongly indicate thatTH9507 has a therapeutic potential in immune indications. In particular,its effect of the T-lymphocyte proliferation response followingvaccination makes it attractive to develop in clinical situations wherethe cell-mediated immune system is depressed, such as viral infectionsin the elderly and immune-deficient states following HIV infection,high-dose chemotherapy or radiotherapy.

Example 4 ThGRF's Benefits in Wasting/Catabolic Conditions

The present example shows the effect of a 7-day administration of TH9507on circulating IGF-1 levels in healthy middle-aged men.

Material and Methods

The study used a randomized, double-blind, placebo-controlled design andwas conducted in healthy men, aged 50 to 60 years old. Subjects (8 pergroup) were injected S.C. once a day for 7 consecutive days withplacebo, 0.5, 1 or 2 mg of TH9507. Circulating IGF-1 levels weremeasured on Days 1 to 7. The 12 hour GH response and TH9507 PK profilewere determined on Day 1 and 7.

Results

As shown in FIG. 5, IGF-1 increased over baseline values by 8%(placebo), 37% (0.5 mg), 89% (1 mg) and 106% (2 mg); these increaseswere statistically significant for all 3 doses of TH9507. The 1 mg and 2mg doses were equally potent and induced a doubling of IGF-1 levels upto levels expected for young adults (286±25 and 284±55 ng/ml,respectively), none of the subjects exhibited levels greater than 400ng/ml. A plateau was reached at Day 4 and 6 for the 1 mg and 2 mg doses,respectively.

GH response to TH9507 increased rapidly both on Days 1 and 2. Theincrease was dose dependent between the 0.5 and 1 mg dose (P<0.01), andwas similar at the 1 mg and 2 mg doses. No significant modification inprolactin, ACTH, cortisol, TSH, LH or FSH was observed following singleor repeated treatment with TH9507.

PK analysis indicated that Cmax and AUC parameters increased in functionof the dose administered. The half-life of the TH9507 ranged between 2and 5 hours.

These results clearly indicate that TH9507 is highly specific on GHsecretion and a powerful IGF-1 inducer, suggesting potential clinicalbenefits in wasting/catabolic conditions.

Example 5 Effects of GRF on Non-HDL Cholesterol in Patients with Type 2Diabetes

The present example illustrates beneficial effects of TH9507 on non-HDLcholesterol levels in a diabetic population.

Material and Methods

A double-blind placebo-controlled study was conducted in 53 type IIdiabetic patients (age=61±7 [SD]; 34% female) on stable antidiabeticmedication (26% on insulin). Patients were randomized to parallel groupsto receive daily subcutaneous administration of a placebo, 1 mg or 2 mgTH9507, respectively.

Results

A statistically significant difference was observed at Week 12 betweenthe 3 treatment groups in the mean total cholesterol change frombaseline (P=0.04). Values decreased in the 2 mg group (−11.1±21.9 mg/dl;−6%), as compared to increases in the Placebo (+9.7±22.6 mg/dl; +5%) and1 mg group (+6.1±16.2 mg/dl; +3%). This effect was accompanied by adecrease in the mean non-HDL cholesterol values in the 2 mg group(−10.1±19.0 mg/dl; −7%) and increases in the placebo (+6.8±17.8 mg/dl;+5%) and 1 mg group (+3.8±15.5 mg/dl; +3%).

No statistically significant differences were observed between the threegroups during the treatment period in terms of insulin relative responseto an oral glucose tolerance test. At Week 12, glycosylated hemoglobin(HbA1c) levels displayed a trend for a decrease in the placebo group, adecrease in the 1 mg group, and no change in the 2 mg group. Clinicallyrelevant changes in antidiabetic medications occurred with a similarincidence in the three treatment groups.

A dose-related increase in the IGF-1 levels was observed at the end ofthe treatment period.

In summary, this study indicates that the repeated administration ofTH9507 for 12 weeks decreases the total and non-HDL cholesterol fractionin diabetic subjects and can be safely administered to this populationwithout impairing glucose control. The effects observed on blood lipidsand the known lipolytic properties of GH warrant the investigation ofTH9507 for the treatment of syndromes associated with visceral fataccumulation.

Example 6 Effect of GRF Analog on Lipid Profile, Body Composition andQuality of Life in HIV-Infected Patients

This study investigated the effects of TH9507, on abdominal fataccumulation associated with HIV lipodystrophy.

Design: Randomized, placebo-controlled, multi-center, dose ranging(placebo, 1 mg or 2 mg SC QD), parallel group, 12-week studyPatients: 61 HIV-infected patients (BMI 28±3 [SD] kg/m², 28% diabetic orglucose intolerant) with increased waist circumference (102 cm±8 [SD])and waist/hip (1.0±0.1 [SD])Measurements: Body composition was assessed by DXA and abdominal CTscan; IGF-I, insulin, glucose, lipid, bone turnover, immunologic, andsafety parameters.Results: Treatment with TH9507 resulted in a dose-related increase inserum IGF-1 levels within the physiological range [18 (32) ng/mL, 87(67) ng/mL, 123 (79) ng/mL, mean (SD) for placebo, 1 mg, 2 mg,respectively, P<0.01 for change in each active group vs. placebo]. Leanbody mass increased significantly in both treatment groups compared toplacebo [−0.5 (1.6) kg, 0.7 (2.0) kg, and 1.7 (2.3) kg; placebo, 1 mg, 2mg, respectively; P<0.01 for 2 mg dose vs. placebo, P<0.05 for the 1 mggroup vs. placebo]. Trunk fat decreased in the 2 mg group compared toplacebo (0.8%, −4.6% and −9.2%; placebo, 1 mg, 2 mg, respectively,P=0.01 for 2 mg vs. placebo), with no significant change in limb fat.Visceral fat decreased most in the 2 mg group (−5.4%, −3.6% and −15.7%;placebo, 1 mg, 2 mg, respectively; P=0.03 for change within 2 mg groupbut P=NS vs. placebo) whereas subcutaneous fat was preserved and did notchange between or within groups. The ratio of VAT:SAT improvedsignificantly in both treatment groups compared to placebo (−0.01(0.10), −0.23 (0.47) and −0.14 (0.18), P<0.01 for the 2 mg group vs.placebo and P<0.05 for the 1 mg group vs. placebo. Triglyceride and thecholesterol to HDL ratio decreased significantly in the 2 mg treatmentgroup compared to placebo. Treatment was generally well-toleratedwithout changes in either fasting glucose or 2-hour OGTT.Conclusion: TH9507 effectively improved body composition and reducedtruncal fat by preferentially decreasing visceral fat while preservingsubcutaneous fat in patients with HIV lipodystrophy. TH9507 did notincrease glucose levels, even among those with impaired glucosetolerance or diabetes mellitus at baseline, and improved the lipidprofile. These data suggest that TH9507, a GRF analog, may be abeneficial treatment strategy for HIV lipodystrophy.

Methods Subjects

Eighty-eight subjects were screened between May 2003 and November 2003.Sixty-one subjects were enrolled. HIV infected males age 18-65 years andnon-menopausal females age >18 years with visceral fat accumulationconsidered to be part of the HIV lipodystrophy syndrome, waistcircumference ≧95 cm for men and ≧94 cm for women, and a waist to hipratio ≧0.94 for men and ≧0.88 for women were enrolled. We excluded,subjects with a BMI ≦20 kg/m2, CD4 cell count ≦100 cells/mm³, viral load≧10,000 copies/mL, history of opportunistic infection or HIV-relateddisease within 3 months of the study, history of prostrate cancer orPSA >5 ng/ml in male subjects, history of breast cancer or abnormalmammography within 6 months of the study (for these patients without amammogram within this time frame, one was performed as part of thestudy) in female subjects, known hypopituitarism or history of pituitarysurgery, radiation or significant head trauma, untreated hypothyroidism,prior history of Type I diabetes mellitus (DM), any prior use of GH orGH related products, experimental or marketed, within 6 months of thestudy, systemic steroid administration or megestrol acetate within 60days of the study, fasting glucose >150 mg/dL, SGOT or SGPT >3× upperlimit of normal, creatinine >1.5× upper limit of normal, or hemoglobin<9 g/dL. Subjects receiving testosterone or estrogen within the priorsix months, with known drug or alcohol dependence or who hadparticipated in another clinical trial with an investigational agentwithin 30 days were also excluded from the study. All subjects wererequired to be on a stable antiretroviral regimen for 8 weeks prior toenrollment. Subjects receiving lipid lowering medications were requiredto be on a stable regimen for 3 months prior to enrollment. Subjectswere not permitted to begin antidiabetic medication, use systemiccorticosteroids for >10 days, or begin estrogen or testosteronepreparations during the study.

Study Procedures

Subjects underwent a screening visit to determine eligibility. Allsubjects gave written consent to participate in the study, and the studywas approved by the Institutional Review Board at each participatingsite. BMI, anthropometic measurements, TSH and prolactin were performed.Viral load and CD4 count were performed if not previously done within 8weeks of the study. A PSA was performed in male subjects and a pregnancytest was performed in female subjects. Mammography was also performed infemale subjects if not previously done within 6 months of the study.

At the baseline visit, anthropometric measurements were obtained. Totalbody lean and fat mass as well as regional fat mass in the trunk andextremities were assessed by DEXA. Abdominal subcutaneous and visceralfat area and the ratio of VAT to SAT were determined by cross-sectionalCT at L4-L5. Biochemical parameters including insulin-like growthfactor-I, glucose, insulin, lipid profile (cholesterol, HDL, LDL,triglyceride, non HDL and the ratio of cholesterol:HDL), free fattyacids, HbA1C, IGFBP-3, and bone markers (serum osteocalcin and NTX-1),CD4, viral load, and a pregnancy test for female subjects weredetermined in the fasting state. In addition, insulin and glucoseresponse to a 75 gram standard glucose challenge were assessed and aquality of life questionnaire was administered.

After baseline evaluations were complete subjects were randomizedequally to one of three groups, placebo, 1 mg TH9507 or 2 mg TH9507.Randomization was performed by an independent consulting firm, using apermuted block algorithm, with block size equal to 3 and keptconfidential in the Theratechnologies Quality Assurance Department untilunblinding. Subjects were stratified for gender at the time of treatmentassignment. Treatment assignment was blinded to investigators andpatients. Active drug and placebo (mannitol) were provided aslyophilized powder in vials of identical appearance, each containing 1.1mg of material. Each vial was reconstituted with sterile waterimmediately prior to injection. Patients were instructed to inject eachAM at approximately 0900 h.

Subjects returned for a safety visit at Week 1 to assess adverse eventsand compliance. Subjects returned at 2 weeks to assess weight,anthropometric measures, lipid profile, free fatty acid, IGFB-1,IGFBP-3, glucose, adverse events and compliance. Subjects returned for avisit at week 6 identical to baseline and a visit at week 9 to assesscompliance and adverse events. Subjects returned for a last study visitat week 12—or, whenever possible, at the time of earlytermination—identical to the baseline visit.

Study Drug

TH9507 was used in this study, as noted above.

Methods Biochemical Indices

Clinical laboratory testing was performed by a central lab. Serum IGF-1was measured after acid-ethanol extraction using the Esoterix RIA kit(Esoterix Inc., Calabasas Hill, Calif.), with a sensitivity of 10 ng/mL.Intra- and inter-assay coefficients of variation were 4.6-20% and 9-10%,respectively. Serum IGFBP-3 was measured using the Esoterix RIA kit(Esoterix Inc., Calabasas Hill, Calif.) with a sensitivity of 0.3 mg/mL.Intra- and inter-assay coefficients of variation were 5.1-13% and5.5-17%, respectively. HDL, LDL, total cholesterol, triglycerides andFFA were determined by enzymatic calorimetric assay. Glucose wasmeasured by enzymatric test (Gluco-quant®, Roche diagnostics,Indianapolis, Ind.). HbA1c was determined by chromatography. PSA wasdetermined using the Hybritech®PSA test (Beckman Coulter, Mississauga,Canada). TSH was measured by microparticle enzyme immunoassay (AxSYMhTSH II, Abbott Laboratories, Abbott Park, Ill.). NTX (N-terminaltelopeptide of type I collagen; a bone resorption marker) was measuredby ELISA using a commercially available kit (Osteomark™, OstexInternational Inc., Seattle, Wash.). Osteocalcin was measured using acommercially available enzymatic immunoassay kit (Metra™ osteocalcin,Quidel Corporation, San Diego, Calif.). CD4 and viral load wereperformed by the individual site labs by routine methodology.

HOMA was calculated as:

$\frac{\left\lbrack \frac{\Delta \mspace{14mu} {Insulin}\mspace{11mu} \left( {{30\mspace{14mu} \min} - {0\mspace{14mu} \min}} \right)}{\Delta \mspace{14mu} {Glucose}\mspace{14mu} \left( {{30\mspace{14mu} \min} - {0\mspace{14mu} \min}} \right)} \right\rbrack}{{Insulin}\mspace{11mu} \left( {0\mspace{14mu} \min} \right)}.$

Body Composition

Whole body and regional DEXA (dual energy X-ray absorptiometry) andcross-sectional abdominal CT scans were performed at the individualstudy sites based on previously established protocols and werestandardized across sites. Digitized images were sent to a centralreading center, St. Lukes Roosevelt Hospital, for analysis byindependent experts without knowledge of treatment assignment. Each sitewas given a procedure manual to follow in obtaining the body compositiondata and was certified in the proper technique. For the CT scans,parameters were standardized as follows: Tube Voltage=120 kV, TubeCurrent=250 mA, Exposure=375 mAs, matrix 512×512. DFOV was set toinclude the entire slice into the field of view. For the DEXA scanstotal body scans were performed with analysis of lean body mass, totalfat mass, and regional fat mass in the trunk and extremities aspreviously described (16). After analysis the data was transferred tothe data management center and incorporated in the study database.

Statistics

Body composition endpoints included trunk fat and trunk to limb fatratio by DEXA and VAT and VAT:SAT by CT scan. Other endpoints includedTGF-I, lipid parameters, glucose and insulin, CD4, viral load, bonemarkers, and quality of life. A sample size of 20 patients/group wasplanned to assess a statistically significant change of 10% or more inintra-group analyses. The planned dropout rate was 25% and therefore 60patients were enrolled into the study. Baseline data are comparedbetween the groups by F-test based on ANOVA for continuous variables andChi-Square test for categorical variables. Change over time betweengroups is compared by ANOVA or the Chi-Square test. Where appropriate,data were rank transformed prior to analysis. Changes within each groupare determined by t-test. The ITT (intent to treat) population isdefined as all subjects who received at least one dose of the studytreatment. Descriptive statistics and analyses for all efficacy andsafety analyses are performed on the ITT population. End of study, 12week, data are used to calculate changes in body composition. Forbiochemical indices, the last observation available is used to calculatechange. Imputation for missing data is not performed. For bodycomposition, end of study data are used to calculate change. Interimanalyses were not performed. Results are mean (SD) unless otherwisenoted. All statistical tests were performed with a two-sided Type Ierror level of 0.05.

Results

Sixty-one subjects were randomized, 21 to placebo, 19 to TH9507 1 mg and21 to TH9507 2 mg. Five subjects discontinued in the placebo group, 2 inthe 1 mg group and 6 in the 2 mg group, for a 79% completion rate. SeeFIG. 6 for patient disposition and flow diagram.

Baseline demographic for the three study groups are shown in Table 3. Atbaseline no significant differences were seen between the groups except,including use of antiretroviral therapy. The percentage of patients withdiabetes (fasting glucose ≧7.0 mmol/L or 2h-glucose ≧11.1 mmol/L) andimpaired glucose tolerance (6.1 mmol/L ≦fasting glucose ≦6.9 mmol/L or7.8 mmol/L ≦2h-glucose <11.1 mmol/L) was not different between thegroups. Among the entire study group, 23% of subjects demonstrated IGTand 5% demonstrated diabetes mellitus at baseline.

TABLE 3 Baseline Characteristics Placebo 1 mg 2 mg P Age 46.1(7.4)46.5(6.4) 44.6(7.1) 0.64 Gender Males/Females 18/3 18/1 18/3 NA Ethnicorigin Cauc./Other 18/3 15/4 17/4 0.84 BMI 28.9(2.7) 28.0(3.2) 27.3(4.1)0.30 WHR  1.0(0.1)  1.0(0.0)  1.0(0.1) 0.65 WC 103.4(8.0)  100.3(6.1) 100.7(9.1)  0.40 IGT-Diabetes 7 4 6 NA Data are mean (SD). BMI,body-mass index; WHR, waist/hip ratio; WC, waist circumference

Body Composition

Lean body mass increased in both treatment groups compared to placebo[−0.5 (1.6) kg, 0.7 (2.0) kg, and 1.7 (2.3) kg, mean (SD) for placebo, 1mg, 2 mg, respectively, P<0.01 for change in 2 mg group vs. placebo,P<0.05 for the change in 1 mg group vs. placebo, Table 3). Total fatdecreased in the 2 mg group compared to placebo [0.3 (1.7) kg, −0.4(1.8) kg and −1.4 (2.0) kg, placebo, 1 mg, 2 mg, respectively, P=0.01for 2 mg group vs. placebo). Trunk fat decreased in the 2 mg groupcompared to placebo (0.8%, −4.6% and −9.2%, placebo, 1 mg, 2 mg,respectively, P=0.01 for 2 mg group vs. placebo) (FIG. 2). VAT tended todecrease more in the 2 mg group (−5.4%, −3.6% and −15.7%, placebo, 1 mg,2 mg, respectively), (P=NS for comparison of 2 mg vs. placebo, P=0.03for change within 2 mg group). SAT did not change significantly betweenthe groups and the ratio of VAT:SAT decreased more in the treatmentgroups compared to placebo [0.01 (0.10), −0.23 (0.47), −0.14 (0.18)](Table II) (P<0.01 for 2 mg vs. placebo and P=0.04 for 1 mg vs.placebo).

Biochemical Indices

IGF-I increased significantly with the 1 and 2 mg dose compared toplacebo [18 (32) ng/mL, 87 (67) ng/mL, 123 (79) ng/mL, placebo, 1 mg 2mg, respectively, P<0.01 for each active group vs. placebo] (FIG. 3).Triglyceride levels decreased in the 2 mg group compared to placebo(−0.2 (1.3), −0.9 (4.2), −0.9 (1.2), last observation values forplacebo, 1 mg, 2 mg, respectively, P<0.05) and the ratio of cholesterolto HDL improved in both treatment groups compared to placebo (0.3 (1.1),−0.3 (0.7), −0.3 (0.6), last observation values for placebo, 1 mg, 2 mg,respectively, P<0.05).

Changes in fasting and 120 minute glucose were not significant betweenor within the treatment groups.

Bone Markers

Osteocalcin increased significantly within the 2 mg group, whereas nochanges with NTX were seen (Table 4).

TABLE 4 Change from Baseline in Body Composition, Biochemical andImmunologic Parameters. Week 12 Placebo 1 mg 2 mg P value P valueBaseline Δ Baseline Δ Baseline Δ 2 mg vs. P 1 mg vs. P Body CompositionLean Body Mass 63.9(9.8)  −0.5(1.6)  64.4(10.0) 0.7(2.0) 65.2(6.9)  1.7(2.3)* 0.0021 0.0467 (kg) Fat Mass (kg) 22.1(6.0)  0.3(1.7)19.0(5.7)  −0.4(1.8)  18.1(7.6)  −1.4(2.0)* 0.0125 0.2962 Trunk Fat (kg)14.6(3.4)  0.1(1.1) 13.1(3.4)  −0.5(1.4)  12.2(4.6)  −1.1(1.3)* 0.01440.1782 Limb Fat (kg) 6.6(2.8) 0.1(0.7) 5.1(2.6) 0.1(0.5) 5.1(3.0)−0.3(0.8)  0.1576 1.0000 VAT (cm²) 190.5(75.7)  −12.0(32.5) 157.8(56.6)  −11.9(28.7)  160.2(53.5)  −21.5(27.9)* 0.6428 1.0000 SAT(cm²) 237.9(85.2)  −10.5(31.6)  188.8(89.0)   6.8(21.2) 189.8(124.3) 4.4(19.8) 0.6919 0.3390 VAT:SAT 0.89(0.50) −0.01(0.10)  1.17(1.25)−0.23(0.47)* 1.12(0.74) −0.14(0.18)* 0.0084 0.0434 Biochemical IndicesGH Parameters¹ IGF-1 (ng/mL) 132.4(42.4)  18.3(31.7) 165.3(62.3)  87.5(66.9)* 157.0(41.4)  122.6(79.1)* 0.0002 0.0042 IGFBP-3 (Mg/L)2.7(0.7) 0.1(0.4) 2.9(0.9)  0.6(0.7)* 2.9(0.4)  0.6(0.5)* 0.0007 0.0022Lipid Parameters¹ NONHDL 4.2(1.0) 0.3(0.8) 4.8(2.1) −0.4(1.5)  4.0(1.0)−0.1(0.7)  0.0770 0.3067 Cholesterol (mmol/L) Total Cholesterol 5.3(1.0)0.4(0.8) 5.9(2.3) −0.4(1.7)  5.2(1.2) −0.1(0.8)  0.1822 0.3175 (mmol/L)LDL (mmol/L) 3.2(0.8)  0.5(0.8)* 3.4(0.9) 0.1(0.8) 3.1(1.0) 0.2(0.7)0.9533 0.3695 HDL (mmol/L) 1.1(0.3) 0.0(0.1) 1.1(0.3) 0.0(0.2) 1.2(0.3) 0.1(0.1)* 1.0000 1.0000 Triglycerides 3.1(2.5) −0.2(1.6)  3.9(4.6)−1.1(4.4)  2.8(1.8) −0.9(1.3)* 0.0131 0.6146 (mEq/L) Chol:HDL 4.9(1.4)0.2(0.8) 5.5(1.3) −0.3(0.6)  4.3(0.7) −0.3(0.7)  0.0128 0.0508 GlucoseParameters Fasting Glucose 5.3(0.6) 0.2(0.8) 5.3(0.8) 0.1(0.5) 5.4(0.6)0.1(0.7) 1.0000 1.0000 (mmol/L) HBA_(1c) (%) 5.1(0.5) −0.3(0.5)*5.2(0.4)  0.2(0.4)* 5.2(0.6) −0.0(0.5)  Contrasts not overall p = 0.0237done(overall p = 0.008) Two hour glucose 6.4(2.9) 0.6(1.8) 6.4(2.4)0.1(2.1) 6.4(2.1) 0.7(1.7) 1.0000 1.0000 (mmol/L) Fasting Insulin15.2(18.0)  5.1(15.8) 22.7(30.7) −2.8(27.6) 12.1(12.5) 7.4(6.2) 1.00000.6799 (mUI/L) HOMA-R 3.7(4.6) 2.1(5.5) 5.5(7.6) 0.8(7.0) 3.0(3.5)2.0(1.6) 1.0000 0.8698 Bone Turnover Markers Osteocalcin (ng/mL)9.2(4.4) 0.5(3.9) 8.0(3.9) 1.5(4.8) 8.3(3.8)  3.0(2.8)* 0.3110 1.0000NTX (nMBCE/mM) 36.3(17.7) −0.8(20.8) 37.1(13.4)  4.5(24.9) 37.2(23.1)−0.5(27.7) 1.0000 1.0000 Immunologic Parameters CD4 (cells/mm³)540.9(252.9)  36.9(107.6) 525.4(309.1)  24.4(147.41)  577.0(327.20)−23.2(70.7)  Contrasts ND overall (overall p = 0.1259 p = 0.3242) Viralload No stats available due to undetectable values ¹For GH and lipidparameters, the p-values presented are obtained from last observationdata. Results are mean (SD) in SI units

Quality of Life

A health-related quality of life questionnaire (PLC, Quality of LifeProfile for the Chronically Ill) was self-administered to 61 patientsrandomized to receive placebo or TH9507 at 1 or 2 mg s.c. daily. The PLCquestionnaire included a general, non-specific part assessing 6dimensions of global health as well as a disease specific part capturingimpact of enlarged abdominal girth, abdominal bloating, tenseness andpain, as well as diarrhea, visible facial changes, visible changes inphysical appearance, and the feeling of being recognized as an HIVpositive person.

Study population included 54 men and 7 women. Baseline mean age was 46±7[SD], BMI 28±3 [SD] kg/m², WC 102 cm±8 [SD] and WHR 1.0±0.1 [SD]. Nosignificant difference between groups was noted for subscales of themain portion of the PLC. Slight changes were observed within the treatedgroups in the positive mood and social well-being scores but were notconsidered clinically significant. Clinically significant improvementswere noted in the enlarged abdominal girth (placebo: −0.13; 1 mg: −0.93,P=0.06 vs baseline; 2 mg: −1.19, P<0.05 vs baseline, NS vs placebo) andbloating scores (placebo: 0.56; 1 mg: −0.50; 2 mg: −0.69, P<0.05 vs.placebo). Improvement in abdominal pain was observed at 1 mg only(P<0.05 vs baseline, NS vs placebo) along with a trend for improvementin tenseness (P=0.07 vs baseline, NS vs placebo). No significant changeswere observed in the other disease-specific items.

These data suggest that administration of TH9507, a GRF analog, in HIVpatients with abdominal fat accumulation improved QOL with regard toenlarged abdominal girth and related items, consistent with the decreasein truncal and visceral fat observed in this population.

AE's and Discontinuation

Discontinuation rates were not different between the groups (24%, 11%,29%, placebo, 1 mg, 2 mg respectively). One subject in the placebo group(arthritis), none in the 1 mg group and 3 in the 2 mg group (rash,arhtralgia, paresthesia) experienced AE's leading to treatmentdiscontinuation. Severe AE's were reported in 6% of the placebo group,13% of the 1 mg group and 10% of the 2 mg group. Musculoskeletal AE'se.g. pain and arthralgias were noted in 24%, 26% and 29% of subjects inthe placebo, 1 mg and 2 mg groups respectively. Carpal tunnel symptomswere not noted in any patient. Edema and/or peripheral swelling werenoted in 1 patient in the 2 mg group only. Headache and/or paresthesiaswere noted in 19%, 32% and 52% of the subjects in the placebo, 1 mg and2 mg groups, respectively. Blood pressure and heart rate did not changebetween or within the groups. One patient in the placebo group comparedto three in the 2 mg group withdrew from the study related to adverseevents. Safety laboratory values, including hemoglobin, WBC, LFT's,creatinine did not differ between the groups (data not shown). CPKincreased in a greater percentage of the 1 (47%) and 2 mg treatedsubjects (38%) compared to placebo (19%), but these changes were smalland did not result in a greater proportion of abnormal CPK valuesbetween the groups. Anti-TH9507 antibodies were not detected after 12weeks in any patient. CD4 count and viral load did not change.

Some of the results using a GRF analog in this study are comparable tothat seen in response to GHRH 1-29 in a recently published study in menwith HIV lipodystrophy, in which truncal fat, but not extremity orsubcutaneous fat decreased in response to physiologic increases in GH(13). The current study extends the findings of Koutkia et al in alarger group of patients, including men and women, using graded doses ofa novel 1-44 amino acid GRF analog that is dosed once rather than twicea day. Although the lower of the 2 doses increased IGF-I significantly,this dose did not result in a significant decrease in trunk fat,suggesting that there may be a threshold increase in IGF-I necessary toreduce truncal fat. Visceral fat also decreased significantly by morethan 15% over 3 months within the 2 mg group. Of note, the magnitude ofthis change on a percentage basis is equivalent to that seen withpharmacologic doses of GH (9), suggesting that this strategy is highlyeffective and potentially very useful because of the general lack ofside effects associated with physiologic increases in GH.

In addition to reducing truncal fat, the 2 mg GRF dose significantlyimproved triglyceride levels and the cholesterol to HDL ratio. This is asignificant advantage of a GRF analog, not seen with other treatmentstrategies for HIV lipodystrophy (17, 18). Similar beneficial effects ontriglyceride were seen with lower, alternating day, but not higher dosesof GH in a study reported by Kotler et al. Growth hormone has been shownto decrease cholesterol and triglyceride levels in GH deficient patientsand among otherwise healthy men chosen for abdominal obesity (19, 20).Taken together, our data suggest that treatment with TH9507 resulted inan improved lipid profile in dyslipidemic, abdominally obese patientswith HIV lipodystrophy.

An important issue regarding the use of GH or related strategies in HIVlipodystrophy is glucose control. Patients with HIV lipodystrophy areoften insulin resistant, and a significant percentage, more than athird, may have impaired glucose tolerance (3). In this study, even withthe higher dose of 2 mg, there were no significant differences infasting glucose or 2-hour glucose in response to a standard GTT and noincrease in HbAIc.

TH9507 was also associated with other benefits in this study.Osteocalcin, a marker of bone formation increased within the 2 mg group,whereas NTX, a marker of bone resorption did not suggesting a netpositive effect on bone turnover. Reduced bone density has beendescribed among patients with HIV disease and among those withlipodystrophy, in inverse association with visceral and truncaladiposity (25, 26). Growth hormone is well known to stimulate boneformation (27). Relative reductions in GH secretion may thereforecontribute to reduced bone density in some patients with lipodystrophyand a positive effect on bone formation, with physiologic increases inGH is an additional benefit of TH9507.

While the invention has been described in connection with specificembodiments thereof, it will be understood that it is capable of furthermodifications and this application is intended to cover any variations,uses, or adaptations of the invention following, in general, theprincipals of the invention and including such departures from thepresent disclosure as come within known or customary practice within theart to which the invention pertains and as may be applied to theessential features hereinbefore set forth, and as follows in the scopeof the appended claims.

REFERENCES

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1-17. (canceled)
 18. A method of altering a first body compositionparameter of a subject, wherein said method results in no orsubstantially no decrease in a second body composition parameter of saidsubject, wherein said second body composition parameter is: (i) limbfat; (ii) subcutaneous fat; (iii) subcutaneous abdominal tissue (SAT);or (iv) any combination of (i) to (iii); said method comprisingadministering to said subject an agent wherein said agent is: (a) agrowth hormone (GH) secretagogue; or (b) a composition comprising a GHsecretagogue and a pharmaceutically acceptable carrier.
 19. The methodof claim 18, wherein said GH secretagogue is GH-releasing factor (GRF)or a GRF analog.
 20. The method of claim 19 wherein said GRF analog is aGRF analog of formula A: X-GRF Peptide (A)

wherein; the GRF peptide is a peptide of formula B;A1-A2-Asp-Ala-Ile-Phe-Thr-A8-Ser-Tyr-Arg-Lys-A13-Leu-A15-Gln-Leu-A18-Ala-Arg-Lys-Leu-Leu-A24-A25- Ile-A27-A28-Arg-A30-R0(B)

wherein, A1 is Tyr or His; A2 is Val or Ala; A8 is Asn or Ser; A13 isVal or Ile; A15 is Ala or Gly; A18 is Ser or Tyr; A24 is Gln or His; A25is Asp or Glu; A27 is Met, Ile or Nle; A28 is Ser or Asn; A30 is a bondor amino acid sequence of 1 up to 15 residues; and R0 is NH₂ orNH—(CH₂)n-CONH₂, with n=1 to 12; and X is a hydrophobic tail anchoredvia an amide bond to the N-terminus of the peptide and the hydrophobictail defining a backbone of 5 to 7 atoms; wherein the backbone can besubstituted by C1-6 alkyl, C3-6 cycloalkyl, or C6-12 aryl and thebackbone comprises at least one rigidifying moiety connected to at leasttwo atoms of the backbone; and said moiety is double bond, triple bond,saturated or unsaturated C3-9 cycloalkyl, or C6-12 aryl.
 21. The methodof claim 20, wherein X is:


22. The method of claim 20, wherein A30 is: (a) a bond; (b) an aminoacid sequence corresponding to positions 30-44 of a natural GRF peptide,or (c) said amino acid sequence of (b) having a 1-14 amino acid deletionfrom its C-terminal.
 23. The method of claim 20, wherein said GRFpeptide is: (a) a polypeptide comprising the amino acid sequence of SEQID NO: 3; (b) a polypeptide comprising the amino acid sequence of SEQ IDNO: 5; or (c) the polypeptide of (a) having a 1 to 14 amino aciddeletion from its C-terminus.
 24. The method of claim 22, wherein saidGRF analog is (hexenoyl trans-3)hGRF(1-44)NH₂.
 25. The method of claim18, wherein said alteration of a first body composition parameter is:(a) an increase in lean body mass (b) a decrease in trunk fat; (c) adecrease in visceral fat; (d) a decrease in abdominal girth; (e) adecrease in visceral abdominal tissue (VAT); (f) a decrease in VAT:SATratio; or (g) any combination of (a) to (f).
 26. The method of claim 18,wherein said first body composition parameter is associated withlipodystrophy, lipohypertrophy, obesity, dyslipidemia,hypertriglyceridemia or syndrome X.
 27. The method of claim 26, whereinsaid lipodystrophy is HIV-related lipodystrophy.
 28. The method of claim18, wherein said subject is HIV positive.
 29. The method of claim 28,wherein said subject is receiving antiviral therapy.
 30. The method ofclaim 18, wherein said subject suffers from diabetes, glucoseintolerance or insulin resistance.
 31. The method of claim 20, whereinsaid GRF analog is administered at a dose of about 0.0001 to 2 mg. 32.The method of claim 31, wherein said GRF analog is administered at adose of about 1 mg or about 2 mg.
 33. The method of claim 18, whereinsaid GH secretagogue or said composition is administered by anintravenous, oral, transdermal, subcutaneous, mucosal, intramuscular,intranasal, intrapulmonary, parenteral, intrarectal or topical route.34. The method of claim 33, wherein said GH secretagogue or saidcomposition is administered by a subcutaneous route.
 35. The method ofclaim 18, wherein said alteration of a first body composition parameterresults in an improvement in quality of life of said subject. 36-88.(canceled)
 89. The method of claim 32, wherein said GRF analog isadministered at a dose of about 2 mg.